Journal: International Journal of Molecular Sciences

Article Title: Decidual Interleukin-22-Producing CD4+ T Cells (Th17/Th0/IL-22+ and Th17/Th2/IL-22+, Th2/IL-22+, Th0/IL-22+), Which Also Produce IL-4, Are Involved in the Success of Pregnancy

doi: 10.3390/ijms20020428

Figure Lengend Snippet: T helper subpopulations present in the decidua and in the peripheral blood of those experiencing unexplained recurrent abortion and those experiencing successful pregnancy. To investigate the CD4+ cell subsets that produce IL-22, the percentages of Th1-, Th2-, Th0-, Th17-, Th17/Th1-, Th17/Th2-, and Th17/Th0 cells, which do not produce IL-22 and which also produce IL-22 (Th1/IL-22+, Th2/IL-22+, Th0/IL-22+, Th17/IL-22+, Th17/Th1/IL-22+, Th17/Th2/IL-22+, and Th17/Th0/IL-22+) were analyzed. Cytokines were measured in the supernatants of the CD4+ T cell clones derived from the decidua and peripheral blood of those experiencing successful pregnancy and those experiencing unexplained recurrent abortion (URA) (Experiment 1 in ) by a multiplex bead-based assay. The statistical analysis was performed with the chi-square test.

Article Snippet: The subpopulations percentages were analyzed by the chi-square test.

Techniques: Clone Assay, Derivative Assay, Multiplex Assay, Bead-based Assay

Journal: International Journal of Molecular Sciences

Article Title: Decidual Interleukin-22-Producing CD4+ T Cells (Th17/Th0/IL-22+ and Th17/Th2/IL-22+, Th2/IL-22+, Th0/IL-22+), Which Also Produce IL-4, Are Involved in the Success of Pregnancy

doi: 10.3390/ijms20020428

Figure Lengend Snippet: CD4+ T subpopulations producing IL-22 in the decidua of those experiencing successful pregnancy and those experiencing URA. The percentage of the CD4+ T cell clones that produce IL-22 in the decidua of those experiencing successful pregnancy and those experiencing unexplained recurrent abortion (URA), which also produce Th2-type (IL-4, IL-5, IL-13), and/or Th1-type (IFN-γ) and/or Th17-type (IL-17A) cytokines was determined. All the cytokines were measured by a multiplex bead-based assay in the supernatant of the CD4+ T cell clones.

Article Snippet: The subpopulations percentages were analyzed by the chi-square test.

Techniques: Clone Assay, Multiplex Assay, Bead-based Assay

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Dynamic motility selection drives population segregation in a bacterial swarm

doi: 10.1073/pnas.1917789117

Figure Lengend Snippet: Spatial segregation of subpopulations with motility heterogeneity in E. coli swarms. (A) Illustration of the protocol to induce motility heterogeneity in E. coli swarms on an antibiotic gradient plate. E. coli YW191 cells (KAN-resistant, labeled by GFP) and YW263 (KAN-sensitive, labeled by Katushka2S) were mixed and inoculated onto the drug-free side of a KAN gradient plate (SI Appendix, Fig. S2 and Methods). The dashed line on the plate marks the boundary between drug-free and drug-infused regions on the plate, and the color scale indicates relative KAN concentration. The spatial distribution of both subpopulations was measured by fluorescence microcopy along the swarm expansion direction (indicated by the black straight arrow) when the swarm had entered the KAN gradient for ∼25 mm. (B) Representative fluorescent image sequence showing the enrichment of the higher-speed subpopulation (YW191, green) near the swarm edge. Red fluorescence was from YW263 cells that had a smaller average speed than YW191 in the drug-infused region of KAN-gradient swarm plates. (C) Representative fluorescent image sequence showing the spatial distribution of YW191 (green) and YW263 (red) cells grown on nonswarming hard agar plates with the same KAN gradient as in B. The image sequences in B and C were taken at different locations whose relative distance to the starting position of the KAN gradient is specified by the ruler below panel C (unit: millimeters; KAN concentration increases from left to right). (Scale bars, 0.1 mm.) (D) Proportion of YW191 cells in swarms on KAN-gradient plates (Left) and in colonies on nonswarming hard agar plates (Middle) plotted against distance to the starting point of the KAN gradient. The population proportion (i.e., ratio between YW191 cell number and total cell number) was measured based on the fluorescence microscopy images as shown in B or C (Methods). The proportion of YW191 cells in swarms on antibiotic-free plates is shown for comparison (Right; distance = 0 mm is located at the plate center). Each line in the plots represents data from an independent colony.

Article Snippet: The population ratio of a subpopulation of cells was computed with a custom-written program in MATLAB based on the background-corrected fluorescence count of the fluorescent images, using the fluorescence count of cells on antibiotic-free plate or on the antibiotic-free side of the gradient plate as the reference.

Techniques: Labeling, Concentration Assay, Fluorescence, Sequencing, Microscopy, Comparison

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Dynamic motility selection drives population segregation in a bacterial swarm

doi: 10.1073/pnas.1917789117

Figure Lengend Snippet: Motion pattern of E. coli swarm cells during the spatial segregation of subpopulations with motility heterogeneity. (A) Representative trajectories of the higher-speed subpopulation (YW191) at ∼5 mm from the swarm edge. The portions of the trajectories moving toward and away from the swarm edge are colored in blue and brown, respectively. (B) Speed distribution of the faster (YW191, green, n = 94) and the slower (YW263, red, n = 314) subpopulations. Lines are Gaussian fits to the speed distributions to obtain the mean and SD of population speed used in main text. (C and D) Angular probability distribution of single-cell velocity directions for the faster (C) and the slower (D) subpopulations, respectively. To generate these plots, single-cell trajectories were divided into 1-s segments and the average velocity direction of these segments was computed as an angle ranging from 0° to 360°, with the swarm expansion direction set as degree 0. The obtained velocity directions were then grouped into 80 polar angle bins of a full circle (360°), with each bin covering an angle of 4.5°. The radii of colored circular sectors in C and D are proportional to the normalized count in the corresponding angle bin and thus represent the probability of single-cell velocity directions falling within the bin. The radius of the dashed circle in each plot indicates a probability of 0.015. (E and F) Average speed of cells plotted against velocity direction for the faster (E) and the slower (F) subpopulations, respectively. In the plots of E and F the polar angle was divided into 80 bins in a way similar to C and D. Single-cell trajectories were divided into 1-s segments and for a specific polar angle bin the average speed of all trajectory segments whose velocity direction fell within this bin was computed. The radii of colored circular sectors in E and F are proportional to the average speed of cells computed for the corresponding polar angle bin, with the radius of the dashed circle indicating a speed of 30 μm/s. Blue and brown colors in C–F indicate moving toward and away from the swarm edge, respectively.

Article Snippet: The population ratio of a subpopulation of cells was computed with a custom-written program in MATLAB based on the background-corrected fluorescence count of the fluorescent images, using the fluorescence count of cells on antibiotic-free plate or on the antibiotic-free side of the gradient plate as the reference.

Techniques: